Extended Nanofluidic Immunochemical Reaction with Femtoliter Sample Volumes
Extended Nanofluidic Immunochemical Reaction with Femtoliter Sample Volumes
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DOI：
10.1002/smll.201302709
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发表时间：
2014-04-01
期刊：
影响因子：
13.3
通讯作者：
Kitamori, Takehiko
中科院分区：
文献类型：
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作者：
Shirai, Kentaro;Mawatari, Kazuma;Kitamori, Takehiko
由于对分析单个细胞的兴趣，对优化免疫测定性能的需求不断增长，导致所需样品量减少。使用超小飞升至阿升样品体积的小型化免疫测定（称为扩展纳米空间的范围）可以满足这种分析需求;然而，在扩展纳米通道中捕获每个靶分子而不损失用于后续检测仍然具有挑战性。这是第一次报告一个成功的扩展的基于纳米流体的定量免疫化学反应，能够高捕获效率，使用一个飞升规模的样品体积。真空紫外低温光刻图形化新方法（100 ℃）键合使得能够在键合之前图案化用于抗体固定的官能团，从而导致仅86 fL的免疫化学反应空间。反应速率分析表明所需样品体积减少至810 fL并且检测限提高至3 zmol，5-6个数量级优于微流体免疫测定形式的可能性。由于纳米级的扩散长度，在秒时间尺度上的高效（接近100%）免疫化学反应是可能的，这对于超低体积样品的分析应该是有利的。
The growing need to optimize immunoassay performance driven by interest in analyzing individual cells has resulted in a decrease in the amount of sample required. Miniaturized immunoassays that use ultra-small femtoliter to attoliter sample volumes, a range known as the extended nanospace, can satisfy this analytical need; however, capturing every targeted molecule without loss in extended nanochannels for subsequent detection remains challenging. This is the first report of a successful extended nanofluidics-based quantitative immunochemical reaction capable of high capture efficiency using a femtoliter-scale sample volume. A novel patterning method using a photolithographic technique with vacuum ultraviolet light and low-temperature (100 degrees C) bonding enables patterning of functional groups for antibody immobilization before bonding, resulting in an immunochemical reaction space of only 86 fL. Reaction rate analyses indicate a decrease in the required sample volume to 810 fL and improvement in the limit of detection to 3 zmol, 5-6 orders of magnitude better than possible with the microfluidic immunoassay format. Highly efficient (near 100%) immunochemical reactions on a seconds time scale are possible due to the nm-scale diffusion length, which should be advantageous for the analysis of ultra-low-volume samples.